发明名称 |
Expression vector for use in a one-step purification protocol |
摘要 |
High expression levels and simple downstream processing are essential for the production of recombinant proteins at low cost. We report here a new expression vector which allows production of fusion proteins in Dictyostelium discoideum. We made use of the ability of Discoidin I to bind Sepharose-4B to set up a nearly single step purification procedure. The Discoidin I coding region was fused to several forms of the malaria parasite CSP gene in a Dictyostelium expression vector, allowing intracellular accumulation as well as partial secretion via a pathway unrelated to the endoplasmic reticulum and Golgi. The fusion proteins present in cell extracts were affinity-purified over Sepharose-4B columns. Addition of a signal peptide allowed endoplasmic reticulum targeting and glycosylation of the fusion protein. Inclusion of a thrombin cleavage site allowed to cleave Discoidin from the CSP protein. The use of stable and low cost Sepharose 4B as affinity matrix should allow large-scale preparations.
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申请公布号 |
US6482607(B1) |
申请公布日期 |
2002.11.19 |
申请号 |
US19990332902 |
申请日期 |
1999.06.15 |
申请人 |
RMF DICTAGENE S.A. |
发明人 |
REYMOND CHRISTOPHE DOMINIQUE;FASEL NICOLAS JOSEPH |
分类号 |
C12N15/09;C07K14/445;C07K14/47;C07K19/00;C12N15/12;C12N15/30;C12N15/62;C12N15/63;C12N15/79;C12P21/02;(IPC1-7):C12N15/63 |
主分类号 |
C12N15/09 |
代理机构 |
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